Autoimmune and inflammatory diseases affect more than fifty million Americans. As a result of basic research in molecular and cellular immunology over the last ten to fifteen years, approaches to diagnosing, treating and preventing these immunological based diseases has been changed forever. By dissecting the individual components of the immune system, those cells, receptors and mediators which are critical to the initiation and progression of immune responses have been, and continue to be, elucidated. Crystallographic analysis of proteins encoded in the major histocompatability complex, identification of an antigen-specific T cell receptor, and development of a basic understanding of the complex cytokine network have all contributed to a revolution in immunology. Equipped with this new and fundamental information about basic immune mechanisms, selective and rational approaches to the treatment of inflammatory and autoimmune disease can now be developed.
Until the last decade, treatment of immunological based disorders were treated exclusively with nonspecific immunosuppressive agents. These included a variety of drugs, such as corticosteroids, antimalarials, methotrexate, azathioprine, and treatments such as total lymphoid irradiation. Although some of these approaches may affect one component of the immune response more than another, they remain nonspecific in their actions and treatment frequently is complicated by serious side effects. It would be very useful to discover and develop new drugs which are immune cell selective or mediator specific and which interfere with processes critical to the initiation, progression, and maintenance of the acute and chronic inflammatory processes associated with certain immunological based diseases.
The two most important cells of the immune response in the autoimmune and inflammatory processes are the T lymphocyte and the monocyte/macrophage.
The T cell is critical to all antigen driven cellular immune responses. There are at least two major subpopulations of T cells: T helper (CD4+) and T cytotoxic (CD8+). T cells recognize antigen via a unique membrane receptor: the T cell antigen receptor (TCR). The TCR can recognize antigen only in association with cell surface proteins known as major histocompatibility complex (MHC) molecules. In response to antigen presented by MHC class II molecules, T helper cells secrete a variety of soluble factors, collectively known as lymphokines. Lymphokines play an essential role in the activation, differentiation, and expansion of all the cells of the immune response. In contrast to the T helper cell, the T cytotoxic cell responds to antigen in the context of MHC class I molecules. Cytotoxic T lymphocytes, once activated, can eliminate cells displaying a specific antigen derived from a virus, tumor cell, or foreign tissue graft.
Mononuclear phagocytic macrophages are widely distributed throughout the body and display great structural and functional heterogeneity. Macrophages are derived from circulating monocytes which migrate into extravascular tissues. The migration of peripheral blood monocytes involves adherence to the endothelium, migration between endothelial cells, and subsequently movement through subendothelial structures. Adherence of monocytes to endothelium involves high molecular weight glycoproteins, such as lymphocyte function-associates antigen 1 (LFA-1; CDlla/CD18), which interacts with intercellular adhesion molecule-1 (ICAM-1; CD54) present on vascular endothelial cells. Monocytes and macrophages produce a variety of pro-inflammatory mediators (cytokines), such as interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor (TNF). These cytokines have numerous effects on many cells within and outside the immune system, such as promoting activation, differentiation, expansion, or apoptosis. In addition, cytokines such as IL-1 increase the expression of adhesion molecules like ICAM-1 and greatly facilitate monocyte migration to the inflammatory site. Furthermore, the monocyte/macrophage is one of the major types of antigen presenting cells required for T helper cell activation.
During the last decade, an understanding of immunopathological reactions has greatly evolved as a result of the characterization of cytokines and interleukins which regulate interactions between cells of the immune system and other nonimmune tissues and cells such as endothelial cells, fibroblasts and adipocytes. A major cytokine increasingly recognized as a central mediator in a wide spectrum of physiologic and immune functions is macrophage-derived Tumor Necrosis Factor-xcex1, also known as TNF-xcex1, or Cachectin. TNF-xcex1 has been found to mediate effects as diverse as tumoricidal activity, wasting and weight loss associated with chronic disease, promotion of cartilage erosion and the destruction of joints in rheumatoid arthritis, and the recruitment of cells to participate more effectively in the host""s response to an invasive agent. In addition, an increasingly large body of evidence indicates that TNF-A serves as the proximal mediator in the evolution of septic shock.
The biological function of TNF-xcex1 extends well beyond its initial discovery as a mediator of tumor necrosis. It is increasingly realized that the interacting milieu of host cytokines existing locally and systemically is an extremely important network that dictates the pathogenesis of many immune and inflammatory diseases. TNF-xcex1 appears to play a critically important role in this regard because of its ability to activate a wide range of cell types in order to promote production of several key cytokines (e.g. IL-1xcex2, IL-1xcex1 and IL-6), bioactive eicosanoids, and platelet activating factor (PAF).
Enhanced synthesis and release of cytokines has been observed during many acute and chronic inflammatory processes, and it is increasingly realized that in many cases, overproduction of TNF-xcex1 is a major contributor to inflammation, cellular injury, and cell death associated with various immunological based diseases.
There is now evidence to indicate that TNF-xcex1 is a primary mediator of septic shock. TNF-xcex1, along with other cytokines, triggers inflammatory and metabolic responses attributed to sepsis and septic shock including adult respiratory distress syndrome (ARDS), fever, and disseminated intravascular coagulation. ARDS is characterized by increased pulmonary capillary permeability resulting in noncardiogenic pulmonary edema, decreased lung compliance and decreased lung volume. Although ARDS is frequently associated with sepsis, it also occurs as a result of smoke inhalation, pancreatitis and long-bone fractures.
Patients infected with the human immunodeficiency virus (HIV) enter a long period of clinical latency prior to developing clinically apparent disease. HIV infects T cells as well as monocytes and macrophages, and activation of latent or marginally active HIV infected cells may be promoted in part by cytokines, including TNF-xcex1. TNF-xcex1 has also been implicated in the pathogenesis of fever, cachexia (wasting syndrome), and Myobacterium tuberculosis infections in patients with acquired immunodeficiency syndrome (AIDS).
Cytokines, including TNF-xcex1, are known to play an important role in the pathogenic processes of inflammatory bowel disease. Ulcerative colitis and Crohn""s disease are two common forms of inflammatory bowel disease.
Complex patterns of interacting cytokines, including TNF-xcex1, and products of arachidonic acid metabolism produced locally in the central nervous system have been implicated in contributing to adverse sequelae of bacterial meningitis.
Rheumatoid arthritis is a heterogenous, systemic disease of unknown etiology, and persons with rheumatoid arthritis typically develop inflammation of joint synovium (synovitis). Clinical symptoms become apparent with progression of synovitis due to production and release of cytokines from activated macrophages along with activation of T lymphocytes, angiogenesis, and attraction of neutrophils to the joint cavities. Cytokines induce synovial cell proliferation, resulting in invasion and destruction of articular cartilage. Synovial fibroblasts are thought to become activated by proinflammatory mediators such as TNF-xcex1 to secrete a large variety of cytokines and growth factors. TNF-xcex1 activity in rheumatoid arthritis includes recruitment and activation of PMNL leukocytes, cellular proliferation, increased prostaglandin and matrix-degrading protease activity, fever, and bone and cartilage resorption. TNF-xcex1 and TNF-xcex1-induced IL-1 induce synthesis of collagenase and stromelysin by synoviocytes, contributing to loss of normal joint integrity and function.
Other diseases/syndromes in which TNF-xcex1 is implicated are vascular injury/atherosclerosis, diabetes mellitus type I, Kawasaki disease, leprosy, multiple sclerosis, anemia of chronic disease, ultraviolet radiation, Helicobacter pylori gastritis/ulcer disease, paracoccidioidomycosis, septic melioidosis, heart failure, familial Mediterranean fever, toxic shock syndrome, chronic fatigue syndrome, allograft rejection, Graft-versus-host disease, Schistosomiasis.
Thus, it would be very useful to provide a means for inhibition of TNF-xcex1 activity in a variety of disease states. The present invention now provides a means for inhibition of TNF-xcex1 activity. This provides a treatment for patients suffering from acute and chronic inflammatory processes associated with various immunological based diseases including septic shock, ARDS, inflammatory bowel disease including ulcerative colitis and Chrohn""s disease, bacterial meningitis, rheumatoid arthritis, fever/cachexia (wasting syndrome)/Myobacterium tuberculosis infections in patients with AIDS, vascular injury/atherosclerosis, diabetes mellitus type I, Kawasaki disease, leprosy, multiple sclerosis, anemia of chronic disease, ultraviolet radiation, Helicobacter pylori gastritis/ulcer disease, paracoccidioidomycosis, septic melioidosis, heart failure, familial Mediterranean fever, toxic shock syndrome, chronic fatigue syndrome, allograft rejection, Graft-versus-host disease, Schistosomiasis. In addition, the present invention provides a treatment which inhibits the activation of latent or marginally active HIV infected cells in patients with AIDS.
The present invention provides compounds having the following general formula (I): 
wherein
Y is nitrogen or CH;
Z1 and Z2 are each independently hydrogen, halogen or NH2; and
A is selected from the group consisting of: 
xe2x80x83wherein X1, X2 and X3 are each independently hydrogen, OH, N3, NH2, N(R)2, NHR, CN, CH2NH2, CONH2, CO2H, CH2OH, SH or SR;
wherein R is C1-C4 alkyl; and
the pharmaceutically acceptable salts thereof;
with the proviso that at least one of X1, X2 or X3 is other than hydrogen and with the further proviso that when Z1 is NH3; Z2 is H or NH2; A is 
xe2x80x83X3 is H or OH; X2 is H; then X1 is not CO2H.
The present invention further provides compounds having the following general formula: 
wherein
Y is nitrogen or CH;
Z1 and Z2 are each independently hydrogen, halogen or NH2; and
X is N3, NH2, N(R)2, NHR, CN, CH2NH2, CONH2, CO2H, CH2OH, SH or SR;
wherein R is C1-C4 alkyl; and
the pharmaceutically acceptable salts thereof.
The present invention further provides a method of inhibiting the TNF-xcex1 activity in a patient in need thereof comprising administering to said patient an effective antiinflammatory amount of a compound of formulas (I) or (II) or of formula (III); 
wherein
Y is nitrogen or CH;
Z1 and Z2 are each independently hydrogen, halogen or NH2; and
the pharmaceutically acceptable salts thereof.
The present invention further provides a method of treating a patient suffering from septic shock comprising administering to said patient an effective immunosuppressant amount of a compound of formulas (I), (II) or (III).
As used herein the term xe2x80x9cC1-C4 alkylxe2x80x9d refers to a saturated straight or branched chain hydrocarbon radical of one to four carbon atoms. Included within the scope of this term are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and the like. The term xe2x80x9chalogenxe2x80x9d or xe2x80x9chaloxe2x80x9d refers to a chlorine, bromine or iodine atom. The term xe2x80x9cPgxe2x80x9d refers to a protecting group such as isopropyldimethylsilyl, tert-butyldiphenylsilyl, methyl-di-tert-butylsilyl, tert-butyldimethylsilyl, benzyl, p-methoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-chlorobenzyl, triphenylmethyl, methoxymethyl, 2-methoxyethoxymethyl, acetate and benzoate. The term xe2x80x9cLgxe2x80x9d refers to a leaving group such as methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, 2-nitrobenzenesulfonate, 3-nitrobenzenesulfonate, 4-nitrobenzenesulfonate or 4-bromobezenesulfonate and the like. It is understood in the art that a protecting group can function as a leaving group and a leaving group can function as a protecting group depending upon the reaction conditions utilized.
The terms xe2x80x9cMsxe2x80x9d or xe2x80x9cmesylatexe2x80x9d refers to a methanesulfonate functionality of the formula: 
The terms xe2x80x9cTsxe2x80x9d or xe2x80x9ctosylatexe2x80x9d refers to a p-toluenesufonate functionality of the formula: 
The term xe2x80x9cpharmaceutically acceptable saltxe2x80x9d refers to those salts that are not substantially toxic at the dosage administered to achieve the desired effect and do not independently possess significant pharmacological activity. The salts included within the scope of this term are hydrobromide, hydrochloride, sulfuric, phosphoric, nitric, formic, acetic, propionic, succinic, glycolic, lactic, malic, tartaric, citric, ascorbic, xcex1-ketoglutaric, glutamic, aspartic, maleic, hydroxymaleic, pyruvic, phenylacetic, benzoic, p-aminobenzoic, anthranilic, p-hydroxybenzoic, salicyclic, hydroxyethanesulfonic, ethylenesulfonic, halobenzenesulfonic, toluenesulfonic, naphthalenesulfonic, methanesulfonic, sulfanilic, and the like. Hydrochloride is preferred as the pharmaceutically acceptable salt of compounds of formulas I, II and III.
It is understood that these compounds of formulas (I), (II) and (III) may exist in a variety of stereoisomeric configurations wherein the substituents X1, X2 and X3 on A of formula (I), substituent X of formula (II) and the hydroxyl on formula (III) may be in the Endo or Exo configuration relative to the bicyclic ring of formula (IV) 
wherein the substituents are as previously defined and A is connected to the ring at the 9-position of formula (IV).
It is further understood that the bicyclic ring compounds defined by A may exist in the CIS or TRANS configuration about the ring juncture. For example the 5xe2x80x945 ring system may have the following configuration 
It is further understood that where the relative configuration is fixed, the maximum number of enantiomers possible for each compound is equal to 2n wherein n represents the total number of chiral centers located on the compound and can be the integer 1, 2, 3 or 4 depending upon the substitution present on the compound. These stereoisomers, including the enantiomers are specifically understood to be included within the scope of the present invention.
The compounds of formula (I) wherein X1, X2 and X3 are each independently H or OH can be prepared as described in Scheme I with the proviso that at least one of X1, X2 or X3 is other than hydrogen. All the substituents, unless otherwise indicated, are previously defined. The reagents and starting materials are readily available to one of ordinary skill in the art. 
Axe2x80x2-OLg is a suitably protected bicyclic sulfonate derivative which is readily available to one of ordinary skill in the art. Axe2x80x2-OLg is selected from the group consisting of: 
wherein X1xe2x80x2, X2xe2x80x2 and X3xe2x80x2 are each independently hydrogen or OPg wherein Pg is a suitable protecting group with the proviso that at least one of X1xe2x80x2, X2xe2x80x2 or X3xe2x80x2 is other than hydrogen. Examples of suitable protecting groups are isopropyldimethylsilyl, tert-butyldiphenylsilyl, methyl-di-tert-butylsilyl, tert-butyldimethylsilyl, benzyl, p-methoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-chlorobenzyl, triphenylmethyl, methoxymethyl, 2-methoxyethoxymethyl, acetate, benzoate and the like. The preferred protecting group is tert-butyldimethylsilyl.
In Scheme I, step A the bicyclic compound of structure (1) is coupled to the suitably protected sulfonate derivative Axe2x80x2-OLg under conditions well known in the art.
For example, the bicyclic compound (1) is combined with a suitable solvent, such as dimethylformamide in a reaction bomb and treated with an equivalent of a suitable base, such as sodium hydride. Examples of appropriately substituted bicyclic compounds (1) are adenine, 2,6-diaminopurine, 6-chloropurine, 2-amino-6-chloropurine, 3-deazoadenine and the like. Adenine is the preferred bicyclic compound (1). The reaction is allowed to stir for about 1 to 3 hours at room temperature to provide the anion of bicyclic compound (1). About 0.3 to 0.4 equivalents of the suitably protected sulfonate derivative Axe2x80x2-OLg is added to the anion of bicyclic compound (1) in the bomb. The bomb is sealed and heated at about 150xc2x0 C. for about 10 to 24 hours. After cooling the product is isolated by extractive techniques well known in the art. For example the reaction is rinsed with water, brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue is then purified by techniques well known in the art, such as flash chromatography on silica gel to provide the protected compound described by structure (2).
In Scheme I, step B the protecting group(s) on protected compound (2) is(are) removed under conditions well known in the art that may vary depending upon the particular protecting group utilized to provide the desired deprotected compound described by formula (Ixe2x80x2) wherein A1 is selected from the group consisting of: 
wherein X1, X2 and X3 are each independently hydrogen or OH with the proviso that at least one of X1, X2 or X3 is other than hydrogen. When more than one protecting group is present on compound (2) they may be removed simultaneously or sequentially depending upon the protecting group employed, the reaction conditions utilized and the product desired by techniques well known and understood in the art of chemistry.
In addition compounds of formula (Ixe2x80x2) can be prepared by treatment of cis-endo-8-hydroxy-bicyclo[3.3.0]octane-endo-2,3-oxirane with the anion of bicyclic compound (1) to provide a mixture of epoxide ring opened products (Iaxe2x80x2) and (Ibxe2x80x2) 
which can be separated by techniques well known in the art such as flash chromatography on silica gel.
The compounds of formulas (I) and (II) wherein X is N3, NHR, N(R)2, CN, SH or SR can be prepared as described in Scheme II. All other substituents, unless otherwise indicated, are previously defined. Starting material for preparation of compounds of formula (II) can be prepared as described in U.S. Pat. No. 4,479,951, Oct. 30, 1984 and U.S. Pat. No. 4,535,158, Aug. 13, 1985. The reagents and starting materials are readily available to one of ordinary skill in the art. 
In Scheme II, step A formula (Ixe2x80x2) or (III) are converted to compound (3) under conditions well known in the art wherein A2 is selected from the group consisting of; 
wherein X1, X2 and X3 are each independently hydrogen or OLg with the proviso that at least one of X1, X2 or X3 is other than hydrogen. Lg is a suitable leaving group. Examples of suitable leaving groups are methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, 2-nitrobenzenesulfonate, 3-nitrobenzenesulfonate, 4-nitrobenzenesulfonate, 4-bromobezenesulfonate and the like. Methanesulfonate is the preferred leaving group.
For example, the appropriately substituted compound of formulas (Ixe2x80x2) or (III) are dissolved in a suitable organic solvent mixture, such as methylene chloride and tetrahydrofuran (5:3). An excess of methanesulfonyl chloride and triethylamine is added and the reaction is stirred for 30 minutes to 3 hours. The reaction is then quenched with water and extracted with a suitable organic solvent, such as methylene chloride. The combined organic extracts are dried over a suitable drying agent, such as anhydrous sodium sulfate, filtered and concentrated under vacuum to provide the compound described by structure (3).
In Scheme II, step B the compounds described by structure (3) undergo a nucleophilic substitution reaction by treatment with a suitable nucleophile to provide the compounds described by formulas (Ixe2x80x3) and (IIxe2x80x2) wherein A3 is selected from the group consisting of: 
wherein X1, X2 and X3 are each independently hydrogen, N3, NHR, N(R)2, CN or SH with the proviso that at least one of X1, X2 or X3 is other than hydrogen.
For example an appropriately substituted compound (3) is dissolved in a suitable solvent. Examples of a suitable solvent are dimethylsulfoxide, dimethylformamide, ethanol and the like. The preferred solvent is ethanol. The solution is then treated with an excess of a suitable nucleophile. Examples of suitable nucleophiles include sodium azide, sodium cyanide, potassium cyanide, lithium cyanide, methylamine, dimethylamine, methyl mercaptide, sodium hydrosulfide, potassium thioacetate and the like. The reaction is stirred at room temperature for approximately 24 hours and then heated at reflux for 2 to 6 hours. Alternatively the reaction can be directly heated at reflux for 2 to 6 hours. The reaction is then concentrated under vacuum and the residue is purified by techniques well known to one skilled in the art. For example, the residue is dissolved in a suitable organic solvent mixture, such as methylene chloride:methanol (9:1) and passed through a plug of silica gel. The filtrate is then concentrated under vacuum to provide the appropriately substituted compound described by formulas (Ixe2x80x3) or (IIxe2x80x2).
The compounds of the formulas (I) and (II) wherein X, X1, X2 and X3 are each independently hydrogen, CH2NH2, CO2H, CONH2 and CH2OH with the proviso that at least one of X, X1, X2 or X3 is other than hydrogen, can be prepared by one of ordinary skill in the art from the corresponding cyano substituted derivative of formulas (Ixe2x80x3) or (IIxe2x80x2), prepared in Scheme II.
For example an appropriately substituted compound of formulas (Ixe2x80x3) or (IIxe2x80x2) wherein X1, X2 and X3 on A3 are each independently hydrogen or CN with the proviso that at least one of X1, X2 or X3 is other than hydrogen, is reduced to the appropriately substituted aminomethyl compound utilizing techniques well known in the art.
For example, the appropriately substituted cyano compound of formulas (Ixe2x80x3) or (IIxe2x80x2) is dissolved in a suitable solvent, such as tetrahydrofuran and treated with an excess of a suitable reducing agent, such as 2M aluminum hydride in tetrahydrofuran. The reaction is refluxed for 2 to 6 hours. Excess reducing agent is carefully decomposed by treatment with acetone and then acidified to pH 7. The mixture is then filtered and the filtrate is concentrated under vacuum. The residue is purified by techniques well known to one skilled in the art. For example, the residue is purified by flash chromatography on silica gel with methylene chloride:methanol (17:3) as eluent to provide the appropriately substituted compounds of formulas (I) and (II) wherein X, X1, X2 and X3 on A are each independently hydrogen or CH2NH2 with the proviso that at least one of X, X1, X2 or X3 is other than hydrogen.
Additionally, an appropriately substituted compound of formulas (Ixe2x80x3) or (IIxe2x80x2) wherein X1, X2 and X3 on A3 are each independently hydrogen or CN with the proviso that at least one of X1, X2 or X3 is other than hydrogen, is hydrolyzed to the appropriately substituted amide derivative utilizing techniques well known in the art.
For example, the appropriately substituted cyano compound of formulas (Ixe2x80x3) or (IIxe2x80x2) is dissolved in a suitable solvent, such as methanol and treated with an equivalent of a suitable base, such as potassium hydroxide. The reaction is heated at reflux for 1 to 5 hours and then concentrated under vacuum. The residue is then purified by techniques well known in the art. For example the residue can be purified by flash chromatography on silica gel utilizing a suitable eluent, such as methylene chloride:methanol to provide the appropriately substituted compounds of formulas (I) and (II) wherein X, X1, X2 and X3 on A are each independently hydrogen or CONH2 with the proviso that at least one of X, X1, X2 or X3 is other than hydrogen.
Additionally, an appropriately substituted compound of formulas (Ixe2x80x3) or (IIxe2x80x2) wherein X1, X2 and X3 on A3 are each independently hydrogen or CN with the proviso that at least one of X1, X2 or X3 is other than hydrogen, is hydrolyzed to the appropriately substituted carboxylic acid derivative utilizing techniques well known in the art.
For example, the appropriately substituted cyano compound of formulas (Ixe2x80x3) or (IIxe2x80x2) is dissolved in a suitable organic solvent, such as tetrahydrofuran. An excess of a suitable base, such as potassium hydroxide is added and the reaction is heated at reflux for approximately 6 hours. After cooling, the reaction is neutralized with a suitable acid, such as 6N hydrochloric acid and the product purified by techniques well known to one skilled in the art. For example, the product can be isolated by ion exchange chromatography to provide the appropriately substituted compounds of formulas (I) and (II) wherein X, X1, X2 and X3 on A are each independently hydrogen or CO2H with the proviso that at least one of X, X1, X2 or X3 is other than hydrogen.
The carboxylic acid derivative described above can then be reduced under conditions well known in the art to provide the corresponding hydroxymethyl derivative.
For example, the appropriately substituted carboxylic acid is dissolved in a suitable organic solvent, such as tetrahydrofuran. An excess of a suitable reducing agent, such as 2M lithium aluminum hydride in tetrahydrofuran is added dropwise to the reaction. The reaction is heated at reflux for 2 to 6 hours. After cooling, excess reducing agent is decomposed by treatment with acetone followed by dilute hydrochloric acid to adjust to pH 7. The mixture is then filtered and the filtrate is concentrated under vacuum. The residue is then purified by techniques well known to one skilled in the art. For example, the residue can be purified by flash chromatography using methylene chloride:methanol (17:3) as the eluent to provide the appropriately substituted compounds of formulas (I) and (II) wherein X, X1, X2 and X3 on A are each independently hydrogen or CH2OH with the proviso that at least one of X, X1, X2 or X3 is other than hydrogen.
The compounds of the formulas (I) and (II) wherein X, X1, X2 and X3 are on A are hydrogen or NH2 with the proviso that at least one of X1, X2 or X3 is other than hydrogen, can be prepared from the corresponding azide derivative [the azide derivative is prepared by nucleophilic substitution of compound (3) with sodium azide as described generally in Scheme II, step B].
For example, the appropriately substituted azide [formulas (Ixe2x80x3) or (IIxe2x80x2) wherein X1, X2, and X3 are each independently hydrogen or N3 with the proviso that at least one of X1, X2 or X3 is other than hydrogen.] is dissolved in a suitable organic solvent, such as tetrahydrofuran and treated with an excess of a suitable reducing agent, such as 2M lithium aluminum hydride in tetrahydrofuran. The reaction is heated at reflux for 2 to 6 hours. After cooling, the excess reducing agent is decomposed with water, the mixture is filtered and the filtrate is concentrated under vacuum. The residue is then purified by techniques well known to one skilled in the art. For example, the residue is purified by flash chromatography using silica gel and a suitable organic eluent, such as methylene chloride:methanol (17:3) to provide the appropriately substituted compounds of formulas (I) and (II) wherein X, X1, X2 and X3 on A are each independently hydrogen or NH2 with the proviso that at least one of X1, X2 or X3 is other than hydrogen.
More specifically the compounds of of formula (I) wherein A is an appropriately substituted octahydropentalene, X1 and X3 are hydrogen, and X2 is OH, can be prepared as described in Scheme III. All the substituents, unless otherwise indicated, are previously defined. The reagents and starting materials are readily available to one of ordinary skill in the art. 
In Scheme III, step A the (+)-diendo-2,6-diacetoxybicyclo[3,3,0]octane (4) [prepared following the procedure described by Henry et al. J. Chem. Soc. Chem. Comm. 1974, 112] is hydrolyzed to the dihydroxy derivative described by structure (5).
For example the (+)-diendo-2,6-diacetoxybicyclo[3,3,0]octane (4) is dissolved in a suitable organic solvent, such as methanol and treated with 2 equivalents of a suitable base, such as potassium hydroxide and allowed to stir at room temperature for 1 to 3 hours. The reaction is then concentrated under vacuum and the residue purified by techniques well known in the art, such as chromatography on silica gel to provide the dihydroxy derivative (5).
In step B the dihydroxy derivative (5) is monoprotected under conditions well known in the art to provide the appropriately substituted monohydroxy derivative described by structure (6).
For example the dihydroxy derivative (5) is dissolved in a suitable organic solvent, such as methylene chloride. It is then treated with a catalytic amount of 4-dimethylaminopyridine, an equivalent of a suitable acid scavenger, such as triethylamine and an equivalent of a suitable protecting group. Examples of suitable protecting groups are tert-butyldimethylsilyl, isopropyldimethylsilyl, methyl-di-tert-butylsilyl, tert-butyldimethylsilyl, acetate, benzoate, tetrahydropyranyl and the like. The preferred protecting group is tert-butyldimethylsilyl. The reaction is allowed to stir for 10 to 24 hours at room temperature. The product is then isolated extractive methods well known in the art. For example the reaction can be washed with water, brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue is then purified by techniques well known in the art, such as flash chromatography on silica gel to provide the monohydroxy derivative (6).
In step C the monohydroxy derivative (6) is converted to the appropriately substituted mesylate described by structure (7) under conditions well known in the art.
For example the monohydroxy derivative (6) is dissolved in a suitable organic solvent, such as methylene chloride. It is then treated with a slight excess of methanesulfonyl chloride and a slight excess of a suitable acid scavenger, such as triethylamine. The reaction is stirred for 1 to 3 hours at room temperature. The mesylate (7) is then isolated by extractive methods well known in the art. For example the reaction is washed with water, brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to provide the mesylate (7).
In step D the mesylate (7) is immediately coupled to the appropriately substituted anion of the bicyclic compound of structure (7a) under conditions that are well known in the art to provide the appropriately substituted compound (8).
For example an appropriately substituted bicyclic compound (7a) is combined with a suitable solvent, such as dimethylformamide in a reaction bomb and then treated with an equivalent of a suitable base, such as sodium hydride. Examples of bicyclic compounds (7a) are adenine, 2,6-diaminopurine, 6-chloropurine, 2-amino-6-chloropurine, 3-deazoadenine and the like. The preferred bicyclic compound (7a) is adenine. The reaction is allowed to stir for about 1 to 3 hours at room temperature to provide the anion of (7a). About 0.3 to 0.4 equivalents of the mesylate (7) are added to the anion of (7a) in the bomb, the bomb is sealed and heated at about 150xc2x0 C. for about 10 to 24 hours. After cooling the product is isolated by extractive techniques well known in the art. For example the reaction is rinsed with water, brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue is then purified by techniques well known in the art, such as flash chromatography on silica gel to provide compound (8).
In step E the protecting group on compound (8) is removed under conditions well known in the art that may vary depending upon the particular protecting group utilized to provide the desired deprotected compound described by structure (9).
For example the appropriately substituted compound (8) wherein Pg is an acid sensitive protecting group such as tert-butyldimethylsilyl group, is dissolved in a suitable solvent mixture, such as methanol and water in about a one to one ratio and then treated with 6N hydrochloric acid until the pH of the reaction is about 2. The reaction is allowed to stir for about 2 to 6 hours and is then concentrated under vacuum to provide the desired deprotected compound (9).
More specifically, the compounds of the formulas (I) and (II) wherein A is an appropriately substituted 5xe2x80x945 bicyclic ring system, X1 and X3 are hydrogen, and X2 or X is CH2NH2, CO2H, CONH2 or CH2OH can be prepared as described in Scheme IV. All other substituents, unless otherwise indicated, are previously defined. The reagents and starting materials are readily available to one of ordinary skill in the art. 
For example, in Scheme IV, step A the cyano derivative described by structure (10) [prepared by nucleophilic substitution of the appropriately substituted derivative of compound (3) with potassium cyanide as described generally in Scheme II, step B] is reduced to the appropriately substituted aminomethyl compound described by structure (11).
For example, an appropriately substituted cyano compound described by structure (10) is dissolved in a suitable solvent, such as tetrahydrofuran and treated with an excess of a suitable reducing agent, such as 2M aluminum hydride in tetrahydrofuran. The reaction is refluxed for 2 to 6 hours. Excess reducing agent is carefully decomposed by treatment with acetone and then acidified to pH 7. The mixture is then filtered and the filtrate is concentrated under vacuum. The residue is purified by techniques well known to one skilled in the art. For example, the residue is purified by flash chromatography on silica gel with methylene chloride:methanol (17:3) as eluent to provide the aminomethyl compound described by structure (11).
In Scheme IV, step B the appropriately substituted cyano compound described by structure (10) is hydrolyzed to the appropriately substituted amide described by structure (12).
For example, an appropriately substituted cyano compound described by structure (10) is dissolved in a suitable solvent, such as methanol and treated with an equivalent of a suitable base, such as potassium hydroxide. The reaction is heated at reflux for 1 to 5 hours and then concentrated under vacuum. The residue is then purified by techniques well known in the art. For example the residue can be purified by flash chromatography on silica gel utilizing a suitable eluent, such as methylene chloride:methanol to provide the purified amide (12).
In Scheme IV, optional step C the appropriately substituted cyano compound described by structure (12) is hydrolyzed to the appropriately substituted carboxylic acid described by structure (13).
For example, an appropriately substituted cyano compound described by structure (10) is dissolved in a suitable organic solvent, such as tetrahydrofuran. An excess of a suitable base, such as potassium hydroxide is added and the reaction is heated at reflux for approximately 6 hours. After cooling, the reaction is neutralized with a suitable acid, such as 6N hydrochloric acid and the product purified by techniques well known to one skilled in the art. For example, the product can be isolated by ion exchange chromatography to provide the carboxylic acid described by structure (13).
In Scheme IV, step D the appropriately substituted carboxylic acid described by structure (13) is reduced to the appropriately substituted alcohol described by structure (14).
For example, an appropriately substituted carboxylic acid described by structure (13) is dissolved in a suitable organic solvent, such as tetrahydrofuran. An excess of a suitable reducing agent, such as 2M lithium aluminum hydride in tetrahydrofuran is added dropwise to the reaction. The reaction is heated at reflux for 2 to 6 hours. After cooling, excess reducing agent is decomposed by treatment with acetone followed by dilute hydrochloric acid to adjust to pH 7. The mixture is then filtered and the filtrate is concentrated under vacuum. The residue is then purified by techniques well known to one skilled in the art. For example, the residue can be purified by flash chromatography using methylene chloride:methanol (17:3) as the eluent to provide the hydroxymethyl derivative described by structure (14).
The compounds of the formulas (I) and (II) wherein A is an appropriately substituted 5xe2x80x945 bicyclic ring system, X1 and X3 are hydrogen, and X or X2 is NH2 can be prepared as described in Scheme V. All other substituents, unless otherwise indicated, are previously defined. The reagents and starting materials are readily available to one of ordinary skill in the art. 
In Scheme V, the appropriately substituted azide [prepared by nucleophilic substitution of sodium azide on compound (3) as described generally in Scheme II, step B] described by structure (15) is reduced to the appropriately substituted primary amine described by structure (16).
For example, an appropriately substituted azide (15) is dissolved in a suitable organic solvent, such as tetrahydrofuran and treated with an excess of a suitable reducing agent, such as 2M lithium aluminum hydride in tetrahydrofuran. The reaction is heated at reflux for 2 to 6 hours. After cooling, the excess reducing agent is decomposed with water, the mixture is filtered and the filtrate is concentrated under vacuum. The residue is then purified by techniques well known to one skilled in the art. For example, the residue is purified by flash chromatography using silica gel and a suitable organic eluent, such as methylene chloride:methanol (17:3) to provide the appropriately substituted primary amine described by structure (16).
The compounds of formula (III) can be prepared following the procedure described by Klessing and Chatterjee in U.S. Pat. No. 4,479,951, Oct. 30, 1984 and by Klessing, K. in U.S. Pat. No. 4,535,158, Aug. 13, 1985 the disclosure of which are hereby incorporated by reference.
The relative configurations encompassed by the stereoisomers of formulas (I), (II) and (III) are readily prepared by one skilled in the art. In addition the enantiomers of formulas (I), (II) and (III) can be resolved utilizing techniques well known in the art of chemistry such as crystallization techniques described by Jacques, J. et al. xe2x80x9cEnantiomers, Racemates, and Resolutionsxe2x80x9d, John Wiley and Sons, Inc., 1981 or by chiral column chromatography.